Association Of Perfluoroalkyl Substances Exposure With The Risk And Severity Of Acute Coronary Syndrome

Part one Determination of plasma perfluoroalkyl substances by ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometryObjective: To develop an economical and efficient approach for the quantification of perfluoroalkyl substances(PFAS)in plasma by employing Ultra-Performance Liquid Chromatography-Quadrupole-Electrostatic Field Orbital Trap High Resolution Mass Spectrometry(UPLC-Q-Orbitrap HRMS).Methods: Chromatographic conditions: Waters ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7 μm)column;The mobile phase consisted of a 5 m M ammonium formate solution and methanol,employing a gradient elution at a flow rate of 0.3 m L·min-1.The column temperature was maintained at 30℃,and the injection volume was 3 μL.Mass spectrometry conditions: HESI electrospray ionisation source,negative ion mode,primary mass spectrometry HRMS full scan plus data-dependent secondary mass spectrometry scan;primary mass spectrometry full scan resolution: 70,000;scan range: 100-800 m/z;secondary mass spectrometry scan resolution: 17500.Plasma sample preparation was carried out using the precipitated protein method.Methodological validation included assessments of selectivity,linearity,lower limit of quantification,limit of detection,precision,accuracy,matrix effects,recovery,and stability.The target analytes included perfluorooctanoic acid,perfluorooctane sulfonic acid,perfluorobutanoic acid,perfluorovaleric acid,perfluorocaproic acid,perfluorogentanoic acid,perfluorononanoic acid,perfluorodecanoic acid,perfluorobutane sulfonic acid,and perfluorocaproic acid.Results: PFAS was well linear in the calibration range,with R2 ranging from 0.990 to 0.999.Under the experimental conditions,the the limits of detection for PFAS were between 0.029 and 0.049 ng·m L-1,and the lower limits of quantification ranged from 0.046 to 0.14 ng·m L-1.The matrix effects for PFAS,expressed as relative standard deviation(RSD)values,were found to be within 1.67% and 6.41%.The accuracy,precision,and stability of the method for PFAS were validated to be in compliance with the requirements for biological sample detection.Summary: The UPLC-Q-Orbitrap HRMS method was established for the determination of PFAS concentrations in human plasma.The method features simplicity of operation,short analysis time,as well as excellent accuracy and precision.It can be effectively utilized for the assessment of PFAS exposure levels in plasma samples.Part two Association of perfluoroalkyl substances exposure and risk of the acute coronary syndromeObjective: To investigate the association between perfluoroalkyl substances exposure and the risk of Acute coronary syndrome(ACS).Methods: This study included 355 newly diagnosed ACS patients,matched(1:1)by sex and age(±5 years)to a control population of 355.Human PFAS exposure levels were measured using the UPLC-Q-Orbitrap HRMS technique.The relationship between PFAS exposure and ACS risk was analyzed using conditional logistic regression.The dose-response relationship between PFAS and ACS risk was analyzed using restricted cubic spline regression modality.The relationship between PFAS mixture exposure and ACS risk was assessed using the quantile g-computation(qgcomp)model by sex(male,female)and age(<50 years,≥50 years)for stratified analyses,with sensitivity analyses performed after excluding age <30 years and BMI >30 kg/m2,respectively.A mediating effect model was applied to assess the mediating effect of platelet parameters between PFAS and ACS risk.Adjusted covariates included age,sex,body mass index(BMI),smoking,alcohol consumption,education level,hypertension,diabetes mellitus and hyperlipidemia.Results: Conditional logistic regression results showed that perfluorooctane sulfonate(PFOS)was significantly and positively associated with the risk of ACS,but not in women.The odds ratio(OR)and 95% confidence interval(95% CI)for PFOS and ACS risk were 1.91(1.22,3.00).Restricted cubic spline results showed a positive increasing linear relationship between PFOA and PFOS and risk of ACS.The qgcomp models showed no significant correlation between PFAS mixture exposure and ACS risk.Mediating effects analysis showed a significant mediating effect of platelet count in the relationship between PFOS and ACS risk.Summary: PFOS exposure is associated with an increased risk of ACS,while no significant association is observed between PFAS mixture exposure and ACS risk.Platelet counts showed a significant mediating effect between PFOS exposure and ACS risk.Part three: Association of perfluoroalkyl substances exposure and the severity of acute coronary syndromesObjective: To investigate the association between perfluoroalkyl substances exposure and the severity of acute coronary syndromes.Methods: 571 patients with newly diagnosed ACS were included in this study.Plasma PFAS exposure levels were measured using the UPLC-Q-Orbitrap HRMS technique.The severity of ACS was assessed through Gensini scores and the number of coronary stenosis.The relationship between PFAS exposure and ACS severity was assessed using an ordered logistic regression model.The dose-response relationships between PFAS exposure and ACS severity were assessed using restricted cubic splines.Threshold effects model was applied to investigate potential threshold effects of PFAS exposure on ACS severity.The qgcomp model was employed to assess the effect of PFAS mixture exposure on ACS severity.Adjusted covariates included age,sex,BMI,smoking,alcohol consumption,education level,hypertension,diabetes,and hyperlipidemia.Results: Ordered logistic regression analysis showed a significant positive association between PFOS and ACS severity,with OR(95% CI)of 1.33(1.10,1.61)and 1.43(1.16,1.76).The restricted cubic spline results showed a positive linear dose-response relationship between PFOS and ACS severity.The threshold effect analysis indicated a significant threshold effect of PFAS on ACS severity.The qgcomp model results showed that PFOS contributed the most to the influence on ACS severity,but no significant impact was observed for PFAS mixture exposure on ACS severity.Summary: A significant positive association is observed between PFOS and the severity of ACS,and a substantial threshold effect exists for PFAS in relation to ACS severity.However,PFAS mixture exposure does not exhibit a significant impact on the severity of ACS.Conclusions:1.PFOS exposure increases the risk of developing ACS.2.Platelet count significantly mediated the association between PFOS exposure and ACS risk.3.PFOS exposure contributes to an increased severity of ACS.4.There was a significant threshold effect between PFAS exposure and ACS severity.5.There was no significant association between PFAS mixture exposure and both ACS risk and severity.